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ID   PWM521     preliminary; circular DNA; SYN; 1993 BP.
XX
AC   M34519;
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DT   27-JUL-1990 (Rel. 5, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pWM521 - complete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-1993
RC   oligos from pWM500 & pWM501 & pWM502 & pWM507
RC   pWM521 from oligo
RC   pWM528 from M13mp18 & pWM521
RC   pWM529 from M13mp19 & pWM521
RA   Mandecki W., Hayden M.A., Shallcross M.A., Stotland E.;
RT   "A totally synthetic plasmid for general cloning, gene expression
RT   and mutagenesis in Escherichia coli";
RL   Gene 94:103-107(1990).
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RN   [2]
RC   pWM521
RA   Mandecki W.;
RT   ;
RL   Submitted (22-MAY-1990) to GenBank on computer-readable media by:
RL   Mandecki W., Abbott Laboratories, Corporate Molecular Biology D93D,
RL   Abbott Park IL 60064.
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RN   [3]
RC   pWM500, pWM501, pWM502, pWM507 from pUC9 & oligo
RA   Mandecki W., Bolling T.J.;
RT   "FokI method of gene synthesis";
RL   Gene 68:101-107(1988).
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RN   [4]
RC   pWM300 from pUC9 & oligo
RA   Mandecki W.;
RT   "Oligonucleotide-directed double strand break repair in plasmids of
RT   Escherichia coli: a method for site-directed mutagenesis";
RL   Proc. Natl. Acad. Sci. U.S.A. 83:7177-7181(1986).
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CC   NCBI gi: 208242
CC   NM (pWM521)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP ()
CC   HO (E.coli)
CC   CP ()
CC   FN (cloning)(mutagenesis)(expression)
CC   SE (color blue/white)
CC   PA (fd)(E.coli)
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. oligo amp gene FokI-FokI
FT                   2. oligo lacZ gene FokI-FokI
FT                   3. oligo ori FokI-FokI
FT                   -> pWM521 1993bp"
FT   mRNA            81..327
FT                   /note="RNA E. coli lacZ gene fragment mRNA"
FT   CDS             119..301
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ)"
FT   misc_binding    136..137
FT                   /note="MCS unique SmaI"
FT   terminator      301..336
FT                   /note="TER E. coli trpA gene terminator"
FT   promoter        337..402
FT                   /note="PRO E. coli bla gene promoter P3"
FT   mRNA            403..1329
FT                   /note="RNA E. coli bla gene mRNA"
FT   CDS             438..1298
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   terminator      1299..1342
FT                   /note="TER bacteriophage fd terminator"
FT   promoter        1343..1408
FT                   /note="PRO E. coli RNAII gene promoter"
FT   mRNA            1409..1962
FT                   /note="RNA E. coli RNAII gene mRNA"
FT   terminator      1415..1440
FT                   /note="TER E. coli RNAI gene terminator"
FT   mRNA            1422..1517
FT                   /note="RNA E. coli RNAI gene mRNA"
FT   promoter        1518..1555
FT                   /note="PRO E. coli RNAI gene promoter"
FT   misc_binding    1961..1962
FT                   /note="SIT E. coli RNaseH cleavage site"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
XX
SQ   Sequence 1993 BP; 524 A; 475 C; 500 G; 494 T; 0 other;
     gaattgatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctg
     ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     gactatgatt acgcccgggc ttgccgtcgt tttacagcga cgagactggg aaaatcctgg
     cgttacccaa cttaatcgcc ttgccgcaca cccccctttc gccagttggc gtaatagcga
     agaagcccgc accgaccgcc cttcccaaca gttgcgtagt ctgaatggcg aatggcgtta
     aactagtagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt ttatttttct
     aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta aaagacgcag
     aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc ggtaaaatcc
     ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa gttctgctat
     gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga cgaatacact
     attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt acggacggaa
     tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact gcggcgaact
     tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac aatatgggag
     accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata ccaaacgacg
     agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta ttaactggcg
     aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg gataaagttg
     caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat aaatctgggg
     caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt aagccttccc
     gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga aatagacaga
     ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt aaaggctcct
     tttggagcct ttttttttgg acggaccgag tagaaaagat caaaggatct tcttgagatc
     ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggacaaagg
     cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     ttttacggtt cct
//